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| ==Procedure==
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| #UVProbe was opened.
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| ##Window > 1. Kinetics
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| ##Methods Icon
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| ###Wavelength: 265nm
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| ###Duration: 300 seconds (5minutes)
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| ###OK
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| #Shimadzu CPS-Controller was set to 25°C.
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| ##wait for the temperature to raise to 25°C
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| #place the sample in the cell and click start.
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| *Phosphate buffer was used as a blank and was used to create the baseline.
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| *Kinetics was run at 265nm using 2.38mL phosphate buffer, 600μL of 500μM stock adenosine, and 20μL ADA (as outlined in the table below).
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| **The velocity was determined by clicking Operations > Activity Table > dAbs. The average of the dAbs values is the average slope (velocity).
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| *5mM adenosine stock solution was too concentrated and exceeded the capacity of the UV-Vis therefore it was diluted:
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| **150 uM adenosine stock solution was used; (stock) 150uL 5mM stock + 850 uL buffer =750 uM ->use .6mL new stock for ADA assay
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| **200 uM adenosine stock solution was used: (stock) 200 uL 5mM stock+800 uL buffer = 1000uM ->use 0.6mL new stock for ADA assay
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| **This table is the volumes of 5mM adenosine and phosphate buffer used to make the new adenosine stock solutions.
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| ***[[Image:Screen_Shot_2013-02-05_at_9.01.20_PM.png]]
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| **This table is the updated table of the volumes and concentrations of solutions that will be added in the cuvette.
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| ***[[Image:Screen_Shot_2013-02-05_at_9.01.33_PM.png]]
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| ==Data== | | ==Data== |
Project name
